Colonoscopy-based colorectal cancer modeling in mice with CRISPR-Cas9 genome editing and organoid transplantation.

Published

Journal Article

Most genetically engineered mouse models (GEMMs) of colorectal cancer are limited by tumor formation in the small intestine, a high tumor burden that limits metastasis, and the need to generate and cross mutant mice. Cell line or organoid transplantation models generally produce tumors in ectopic locations-such as the subcutaneous space, kidney capsule, or cecal wall-that do not reflect the native stromal environment of the colon mucosa. Here, we describe detailed protocols to rapidly and efficiently induce site-directed tumors in the distal colon of mice that are based on colonoscopy-guided mucosal injection. These techniques can be adapted to deliver viral vectors carrying Cre recombinase, CRISPR-Cas9 components, CRISPR-engineered mouse tumor organoids, or human cancer organoids to mice to model the adenoma-carcinoma-metastasis sequence of tumor progression. The colonoscopy injection procedure takes ∼15 min, including preparation. In our experience, anyone with reasonable hand-eye coordination can become proficient with mouse colonoscopy and mucosal injection with a few hours of practice. These approaches are ideal for a wide range of applications, including assessment of gene function in tumorigenesis, examination of tumor-stroma interactions, studies of cancer metastasis, and translational research with patient-derived cancers.

Full Text

Duke Authors

Cited Authors

  • Roper, J; Tammela, T; Akkad, A; Almeqdadi, M; Santos, SB; Jacks, T; Yilmaz, ÖH

Published Date

  • February 2018

Published In

Volume / Issue

  • 13 / 2

Start / End Page

  • 217 - 234

PubMed ID

  • 29300388

Pubmed Central ID

  • 29300388

Electronic International Standard Serial Number (EISSN)

  • 1750-2799

Digital Object Identifier (DOI)

  • 10.1038/nprot.2017.136

Language

  • eng

Conference Location

  • England